Design and Use of Organic Voltage Sensitive Dyes
The chemistry and the physics of voltage sensitive dyes (VSDs) should be understood and appreciated as a prerequisite for their optimal application to problems in neuroscience. This chapter provides a basic understanding of the properties of the large variety of available organic VSDs. The mechanisms by which the dyes respond to voltage guides the best setup of the optics for recording or imaging electrophysiological activity. The physical and chemical properties of the dyes can be tuned to optimize delivery to and staining of the cells in different experimental preparations. The aim of this chapter is to arm the experimentalists who use the dyes with enough information and data to be able to intelligently choose the best dye for their specific requirements.
KeywordsFluorescence Resonance Energy Transfer Second Harmonic Generation Molecular Orbital Calculation Membrane Potential Change Voltage Sensitivity
I am indebted to the many talented chemists, microscopists, and neuroscientists who have collaborated with me and who have carried out much of the research summarized in this chapter. Most notably, I wish to acknowledge my long-term collaborators Larry Cohen, Aaron Lewis, Mei-de Wei, and Joe Wuskell. The more recent work in my lab has benefited from collaborations with Ping Yan and Srdjan Antic. This work was supported by NIH EB001963.
- Beach JM, McGahren ED, Xia J, Duling BR (1996) Ratiometric measurement of endothelial depolarization in arterioles with a potential-sensitive dye. Am J Physiol 270:2216–2227.Google Scholar
- Loew LM (1993) Confocal microscopy of potentiometric fluorescent dyes. Methods Cell Biol 38:194–209.Google Scholar
- Loew LM (1994) Voltage sensitive dyes and imaging neuronal activity. Neuroprotocols 5:72–79.Google Scholar
- Loew LM (2001) Mechanisms and principles of voltage sensitive fluorescence. In Rosenbaum DS, Jalife J (eds) Optical mapping of cardiac excitation and arrhythmias. Futura Publishing, Armonk.Google Scholar
- Millard AC, Campagnola PJ, Mohler W, Lewis A, Loew LM (2003a) Second harmonic imaging microscopy. In Marriott G, Parker I (eds) Methods in enzymology, vol 361B. Academic Press, San Diego.Google Scholar
- Millard AC, Lewis A, Loew LM (2005b) Second harmonic imaging of membrane potential. In imaging in neuroscience and development. In: Yuste R, Lanni F, Konnerth A (eds) Imaging neurons a laboratory manual. Cold Spring Harbour Laboratory Press, New York.Google Scholar
- Ross WN, Salzberg BM et al (1977) Changes in absorption, fluorescence, dichroism, and birefringence in stained giant axons: optical measurement of membrane potential. J Membr Biol 3:141–183.Google Scholar